Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

Monika Taucher; Kathrin Breuker

doi:10.1016/j.jasms.2010.02.025

Top-down mass spectrometry for sequencing of larger (up to 61 nt) RNA by CAD and EDD

J Am Soc Mass Spectrom. 2010 Jun;21(6):918-29. doi: 10.1016/j.jasms.2010.02.025. Epub 2010 Mar 9.

Authors

Monika Taucher¹, Kathrin Breuker

Affiliation

¹ Institute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, Innsbruck, Austria.

PMID: 20363646
DOI: 10.1016/j.jasms.2010.02.025

Abstract

We have studied the effect of solution additives on hydrolysis and charge state distribution in ESI MS of RNA. Lower and higher charge state ions can be electrosprayed from solutions containing 25 mM piperidine/25 mM imidazole and 1% vol. triethylamine, respectively, with base-catalyzed hydrolysis rates that are sufficiently slow to perform MS/MS experiments. These lower and higher charge state ions are suitable as precursors for CAD and EDD, respectively. We demonstrate nearly complete sequence coverage for 61 nt RNA dissociated by CAD, and 34 nt RNA dissociated by EDD, and suggest a mechanism for backbone fragmentation in EDD of RNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Mass Spectrometry / methods*
Molecular Sequence Data
RNA / chemistry*

Substances

RNA

Grants and funding

Y 372/FWF_/Austrian Science Fund FWF/Austria